This study aimed to explore the effect of FM substitution by defatted Tenebrio molitor larvae meal (dTM) on the response of European seabass to an acute stress challenge. An FM-based diet was used as a control and two other isoproteic/isoenergetic diets were formulated to replace 50 and 100% of FM by dTM. Each diet was tested in quadruplicate groups of 15 fish (69 ± 5 g) fed until visual satiety for 16 weeks. After the feeding trial, fish were subjected to 1 min air exposure followed by 1 h of recovery before sampling. The haematological profile, plasma metabolites, and humoral immunity biomarkers, as well as hepatic oxidative stress and antioxidant capacity, were analysed. A clear response to acute stress was observed by a significant increase in haemoglobin, haematocrit, red blood cells, and almost all evaluated plasma metabolites and humoral parameters, regardless of dietary treatment. The obtained results demonstrated that partial substitution of FM by IM did not affect the stress response of seabass. However, total FM replacement increased the hepatic activity of total peroxidase and superoxide dismutase in fish fed TM100.